LOCUS       sequence-190839-        5427 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP tagged zebrafish ASC in the pCS2+ vector for in vitro 
            transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5427)
  AUTHORS   Vincent WJ, Freisinger CM, Lam PY, Huttenlocher A, Sauer JD
  TITLE     Macrophages mediate flagellin induced inflammasome activation and 
            host defense in zebrafish.
  JOURNAL   Cell Microbiol. 2016 Apr;18(4):591-604. doi: 10.1111/cmi.12536. Epub
            2015 Nov 4.
  PUBMED    26468080
REFERENCE   2  (bases 1 to 5427)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1111/cmi.12536";
            journalName: "Cell Microbiol"; date: "2016-04"; volume: "18"; issue:
            "4"; pages: "591-604"
COMMENT     Sequence Label: pCS2+ gfp-zASC
FEATURES             Location/Qualifiers
     source          1..5427
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..43
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     25..42
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      82..91
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             88..804
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    1469..1603
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1493..1512)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1547..1566
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1716..1736)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1716..1734)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1755..1771)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1755..1771)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1768..1790)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1779..1795
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1803..1809,1810..1827,1828..1833))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1848..1869
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1986..2003)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2157..2745)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2237..2256)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2916..3707,3708..3776))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3539..3558
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3777..3881)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3907..4362)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4044..4065)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4256..4275
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4433..5417
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag ctacttgttc
       61 tttttgcagg atccaccggt cgccaccatg gtgagcaagg gcgccgagct gttcaccggc
      121 atcgtgccca tcctgatcga gctgaatggc gatgtgaatg gccacaagtt cagcgtgagc
      181 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
      241 ggcaagctgc ctgtgccctg gcccaccctg gtgaccaccc tgagctacgg cgtgcagtgc
      301 ttctcacgct accccgatca catgaagcag cacgacttct tcaagagcgc catgcctgag
      361 ggctacatcc aggagcgcac catcttcttc gaggatgacg gcaactacaa gtcgcgcgcc
      421 gaggtgaagt tcgagggcga taccctggtg aatcgcatcg agctgaccgg caccgatttc
      481 aaggaggatg gcaacatcct gggcaataag atggagtaca actacaacgc ccacaatgtg
      541 tacatcatga ccgacaaggc caagaatggc atcaaggtga acttcaagat ccgccacaac
      601 atcgaggatg gcagcgtgca gctggccgac cactaccagc agaatacccc catcggcgat
      661 ggccctgtgc tgctgcccga taaccactac ctgtccaccc agagcgccct gtccaaggac
      721 cccaacgaga agcgcgatca catgatctac ttcggcttcg tgaccgccgc cgccatcacc
      781 cacggcatgg atgagctgta caagaattca gatctcaaga tggcggaatc tttcaaggag
      841 cacctgcagg aggcttttga ggatctgggt gctgataatc ttagaaagtt taaaagcaaa
      901 ctgggcgatc ggaggcagga accgcgcgtc acaaagtctg caatcgaaaa gctgaaagac
      961 gagatcgatc tggctgattt aatggtcggc gtgttcacat caaaagacgc ggtttctgta
     1021 acagtggaga ttttgagggc gatcaagtgt aatgcagtag cagatgatct attgaggaac
     1081 acagggcaat cagaaagcaa aggtgctccc tctgatgaaa gcaaatgtgc ttcctctaag
     1141 gctgtaagca aagttgcttt ctccaaggta aacttcatcg atgagcactg gaaagaattg
     1201 atagaccgag ttaataatgt ggaccccatc ctagatatac ttcggcagaa aaaagtcatc
     1261 acaaatgagg attactgtac catccgtaat aaggagactc cccaaaagaa gatgagagag
     1321 ttattaacag gcccaatcac atgtgctggc aacaaaggga aagaagtctt gtatgatgct
     1381 ctcagagaga gcaataaatt cctgatggat gaccttgagg atgctcagtg actcgagcct
     1441 ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca ttgatgagtt
     1501 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     1561 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     1621 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg cggccgcggc
     1681 gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc
     1741 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     1801 cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     1861 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     1921 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     1981 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2041 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2101 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2161 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2221 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2281 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2341 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2401 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2461 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2521 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2581 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2641 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2701 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     2761 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     2821 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     2881 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     2941 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3001 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3061 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3121 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3181 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3241 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3301 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3361 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3421 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     3481 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     3541 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     3601 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     3661 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     3721 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     3781 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     3841 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     3901 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3961 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     4021 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     4081 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     4141 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     4201 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     4261 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     4321 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     4381 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagtcgaccg
     4441 ccaattcaat atggcgtata tggactcatg ccaattcaat atggtggatc tggacctgtg
     4501 ccaattcaat atggcgtata tggactcgtg ccaattcaat atggtggatc tggaccccag
     4561 ccaattcaat atggcggact tggcaccatg ccaattcaat atggcggacc tggcactgtg
     4621 ccaactgggg aggggtctac ttggcacggt gccaagtttg aggaggggtc ttggccctgt
     4681 gccaagtccg ccatattgaa ttggcatggt gccaataatg gcggccatat tggctatatg
     4741 ccaggatcaa tatataggca atatccaata tggccctatg ccaatatggc tattggccag
     4801 gttcaatact atgtattggc cctatgccat atagtattcc atatatgggt tttcctattg
     4861 acgtagatag cccctcccaa tgggcggtcc catataccat atatggggct tcctaatacc
     4921 gcccatagcc actcccccat tgacgtcaat ggtctctata tatggtcttt cctattgacg
     4981 tcatatgggc ggtcctattg acgtatatgg cgcctccccc attgacgtca attacggtaa
     5041 atggcccgcc tggctcaatg cccattgacg tcaataggac cacccaccat tgacgtcaat
     5101 gggatggctc attgcccatt catatccgtt ctcacgcccc ctattgacgt caatgacggt
     5161 aaatggccca cttggcagta catcaatatc tattaatagt aacttggcaa gtacattact
     5221 attggaagta cgccagggta cattggcagt actcccattg acgtcaatgg cggtaaatgg
     5281 cccgcgatgg ctgccaagta catccccatt gacgtcaatg gggaggggca atgacgcaaa
     5341 tgggcgttcc attgacgtaa atgggcggta ggcgtgccta atgggaggtc tatataagca
     5401 atgctcgttt agggaaccgc cattctg
//
